皖南医学院学报
皖南醫學院學報
환남의학원학보
ACTA ACADEMIAE MEDICINAE WANNAN
2014年
1期
12-15
,共4页
hsa-miR-30a%骨肉瘤%细胞凋亡%抑癌基因
hsa-miR-30a%骨肉瘤%細胞凋亡%抑癌基因
hsa-miR-30a%골육류%세포조망%억암기인
hsa-miR-30a%osteosarcoma%apoptosis%tumor suppressor gene
目的：研究miR-30a在人骨肉瘤细胞U2-OS中的过表达对其凋亡的影响。方法：构建pEZX-MR04-pre-miR-30a真核表达载体，瞬时转染人骨肉瘤细胞株U2-OS，荧光显微镜及荧光定量PCR法（ qRT-PCR）检测miRNA-30a在U2-OS中的表达，并采用流式细胞仪检测稳转株细胞的凋亡情况。结果：荧光显微镜下观察真核表达载体pEZX M-R04-pre-miR-30a转染后的U2-OS细胞，可见大量绿色荧光；qRT-PCR检测U2-OS中miR-30a的表达明显提高；流式细胞检测转染后的U2-OS细胞的凋亡明显增加。结论：miR-30a在人骨肉瘤细胞U2-OS过表达能促进其凋亡，miR-30a可能是U2-OS细胞潜在的抑癌基因。
目的：研究miR-30a在人骨肉瘤細胞U2-OS中的過錶達對其凋亡的影響。方法：構建pEZX-MR04-pre-miR-30a真覈錶達載體，瞬時轉染人骨肉瘤細胞株U2-OS，熒光顯微鏡及熒光定量PCR法（ qRT-PCR）檢測miRNA-30a在U2-OS中的錶達，併採用流式細胞儀檢測穩轉株細胞的凋亡情況。結果：熒光顯微鏡下觀察真覈錶達載體pEZX M-R04-pre-miR-30a轉染後的U2-OS細胞，可見大量綠色熒光；qRT-PCR檢測U2-OS中miR-30a的錶達明顯提高；流式細胞檢測轉染後的U2-OS細胞的凋亡明顯增加。結論：miR-30a在人骨肉瘤細胞U2-OS過錶達能促進其凋亡，miR-30a可能是U2-OS細胞潛在的抑癌基因。
목적：연구miR-30a재인골육류세포U2-OS중적과표체대기조망적영향。방법：구건pEZX-MR04-pre-miR-30a진핵표체재체，순시전염인골육류세포주U2-OS，형광현미경급형광정량PCR법（ qRT-PCR）검측miRNA-30a재U2-OS중적표체，병채용류식세포의검측은전주세포적조망정황。결과：형광현미경하관찰진핵표체재체pEZX M-R04-pre-miR-30a전염후적U2-OS세포，가견대량록색형광；qRT-PCR검측U2-OS중miR-30a적표체명현제고；류식세포검측전염후적U2-OS세포적조망명현증가。결론：miR-30a재인골육류세포U2-OS과표체능촉진기조망，miR-30a가능시U2-OS세포잠재적억암기인。
Objective:To determine the expression of miR-30a in human osterosarcoma cell line U2-OS and the effects on the apoptosis of human U2-OS cells.Methods:pEZX-MR04-pre-miR-30a vector was constructed and transiently transfected into U2-OS cells.The expression of miRNA-30a in U2-OS cells was verified by fluorescence microscopy and quantitative real time PCR (qRT-PCR).Flow cytometry was used to detect the effects of miR-30a on the apoptosis of U2-OS cells.Results:pEZX-MR04-pre-miR-30a vector was successfully transfected into U2-OS cells.Quantitative green positive cells were seen under fluorescence microscope,and qRT-PCR results indicated significant increase of miR-30a expression in U2-OS cell lines.Flow cytometry findings showed that miR-30a significantly promoted U2-OS cells apoptosis.Conclusion:Over-expression of miR-30a has significantly facilitated the apoptosis cells U2-OS,suggesting that miR-30a may be a potential tumor suppressor gene for human osteosarcoma.
